The proposed research addresses the gap in knowledge in the fundamental control mechanism of genetic transcription of the albumin gene in rat liver. Nonhistone chromosomal proteins that can interact with specific loci in the cloned, rat liver serum albumin gene will be isolated and their functional significance assayed. Isolation will involve mapping and sequencing the high-affinity protein binding sites that are 5', 3' and within the serum albumin gene by the "footprinting" technique. The sequenced site will be tandemly linked and native DNA affinity resins made by covalently attaching DNA to agarous beads for affinity chromatography of a nonhistone protein extract. The high affinity binding nonhistone proteins can be characterized by two dimensional gel electrophoresis stained with silver complexes. The sensitivity of this assay can recognize in the order of 10 molecules of protein/nuclei which extends our investigation to the minor polypeptides of the rat nucleus. The functional significance can be assayed by measuring the levels of the DNA binding nonhistone protein in the nuclei of rat liver under varying intakes of carbohydrate, protein and fat. The levels of protein can be measured with defined DNA substrates retained on nitrocellulose filters in the presence of the protein component. The levels of the protein can be correlated with levels of the nuclear and cytoplasmic transcript by appropriate hybridization probes made from the albumin gene.